title: Brain tumor stem cell multipotency correlates with nanog expression and extent of passaging in human glioblastoma xenografts.
abstract: Glioblastoma multiforme (GBM) is the most common primary brain tumor, with a median survival of only 15 months. A subpopulation of cells, the brain tumor stem cells (BTSCs), may be responsible for the malignancy of this disease. Xenografts have proven to be a robust model of human BTSCs, but the effects of long-term passaging have yet to be determined. Here we present a study detailing changes in BTSC multipotency, invasive migration, and proliferation after serial passaging of human GBM xenografts. Immunocytochemistry and tumorsphere formation assays demonstrated the presence of BTSCs in both early generation (EG-BTSCs; less than 15 passages) and late generation (LG-BTSCs; more than 24 passages) xenografts. The EG-BTSCs upregulated expression of lineage markers for neurons and oligodendrocytes upon differentiation, indicating multipotency. In contrast, the LG-BTSCs were restricted to an astrocytic differentiation. Quantitative migration and proliferation assays showed that EG-BTSCs are more migratory and proliferative than LG-BTSCs. However, both populations respond similarly to the chemokine SDF-1 by increasing invasive migration. These differences between the EG- and LG-BTSCs were correlated with a significant decrease in nanog expression as determined by qRT-PCR. Mice implanted intracranially with EG-BTSCs showed shorter survival when compared to LG-BTSCs. Moreover, differentiation prior to implantation of EG-BTSCs, but not LG-BTSCs, led to increased survival. Thus, nanog may identify multipotent BTSCs. Furthermore, limited passaging of xenografts preserves these multipotent BTSCs, which may be an essential underlying feature of GBM lethality.
authors.affiliation: Medical Scientist Training Program, Mayo Clinic: College of Medicine, Rochester, Minnesota, USA.
authors.forename: Dominique M
authors.initials: DM
authors.lastname: Higgins
authors.forename: Ruisi
authors.initials: R
authors.lastname: Wang
authors.forename: Brian
authors.initials: B
authors.lastname: Milligan
authors.forename: Mark
authors.initials: M
authors.lastname: Schroeder
authors.forename: Brett
authors.initials: B
authors.lastname: Carlson
authors.forename: Jenny
authors.initials: J
authors.lastname: Pokorny
authors.forename: Samuel H
authors.initials: SH
authors.lastname: Cheshier
authors.forename: Fredric B
authors.initials: FB
authors.lastname: Meyer
authors.forename: Irving L
authors.initials: IL
authors.lastname: Weissman
authors.forename: Jann N
authors.initials: JN
authors.lastname: Sarkaria
authors.forename: John R
authors.initials: JR
authors.lastname: Henley
mesh_terms: D000818:Animals; D020295:Brain Stem Neoplasms / Q000378:metabolism*; D002454:Cell Differentiation; D002465:Cell Movement; D049109:Cell Proliferation; D054377:Chemokine CXCL12 / Q000378:metabolism; D005909:Glioblastoma / Q000378:metabolism*; D018398:Homeodomain Proteins / Q000096:biosynthesis / Q000378:metabolism*; D006801:Humans; D051379:Mice; D008819:Mice, Nude; D039902:Multipotent Stem Cells / Q000378:metabolism*; D000071317:Nanog Homeobox Protein; D009361:Neoplasm Invasiveness; D009368:Neoplasm Transplantation; D014411:Neoplastic Stem Cells / Q000378:metabolism*
chemical_list: C515255:CXCL12 protein, human; D054377:Chemokine CXCL12; D018398:Homeodomain Proteins; C486780:NANOG protein, human; D000071317:Nanog Homeobox Protein